Human Cell Viability Assay?

[Robert M. Straubinger]

In article <28SEP93.20645568.0063 at VM1.MCGILL.CA> CX74 <CX74 at MUSICA.MCGILL.CA> writes:
>Does anyone out there have a good, reproducible cell viability
>assay for 96-microwell plates?  I'm trying to measure cellular
>growth and survival in a human cell culture.  If anyone has hints on
>getting the MTT/acid-isopropanol assay to produce reproducible
>results (and perhaps get rid of the percipitate at the bottom
>of the wells) it would be appreciated as well.

  Hmmm. the isopropanol should dissolve those ppt's, if they are MTT.
With the small wells in (say) 96-well plates, it's difficult to agitate
enough to get good isopropanol mixing, and the vibrator in (our) plate
reader doesn't seem to be enough either. Thus we use a multichannel pipettor
and mix all wells vigorously. With variable dissolution of MTT ppt.,
variability can be understandably high.
  Sigma advertises an 'XTT' assay and claims the metabolic product
is soluble. No experience with it here, though.


Bob Straubinger
Pharmaceutics
SUNY Buffalo